研究目的
Investigating the capability of a high-density micro-LED array to achieve precise optogenetic control of intracellular Ca2+ dynamics at the single-cell level in dense cell populations.
研究成果
The high-density micro-LED array demonstrated precise optogenetic control of Ca2+ signaling at the single-cell level, with the ability to address individual cells sub-10 μm apart. This suggests its potential as a lab-on-a-chip platform for single-cell optogenetics, enabling pharmaceutical screening and fundamental studies on cell networks.
研究不足
The energy conversion efficiency of the LED pixels suffered from voltage drop across contact wires. The spatial resolution is limited by the omnidirectional emission from LEDs. The study was conducted at room temperature, potentially affecting cell activity over time.
1：Experimental Design and Method Selection:
The study employed a GaN-based, 4-by-4 micro-LED array for optogenetic stimulation of HEK 293 cells co-expressed with ChR2 and either jRCaMP1a or NIR-GECO1 Ca2+ indicators. The array was designed to output bright, localized, and fast-switching light for precise optogenetic control.
2：Sample Selection and Data Sources:
HEK 293 cells were cultured on PDMS pieces and transfected with optogenetic actuators and Ca2+ indicators. Fluorescence microscopy was used to monitor intracellular Ca2+ dynamics.
3：List of Experimental Equipment and Materials:
A micro-LED array, fluorescence microscope, PDMS pieces, HEK 293 cells, optogenetic actuators (ChR2), and Ca2+ indicators (jRCaMP1a and NIR-GECO1).
4：1).
Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Cells were optogenetically stimulated by LED pixels while monitoring Ca2+ dynamics. Control experiments were conducted to validate the specificity of the optogenetic activation.
5：Data Analysis Methods:
The change in fluorescence intensity (?F/F0) was used to quantify intracellular Ca2+ dynamics. Statistical analysis was performed to assess the significance of the results.
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